Leishmania Amastigotes Induced Remission of Cutaneous Leishmaniasis, Psoriasis, Psoriatic Arthritis and Related Diseases.

Amastigotes from L(L) amazonensis (La); L(L) venezuelensis (Lv); L(V) brasiliensis (Lb) and L(L)chagasi(Lch) were cultured axenically in a liquid culture medium (O’Daly’s medium). Patients from a cutaneous leishmaniasis (CL) endemic and hyperendemic regions in Venezuela, receiving different treatments were followed up for 6 years. Remission of lesions (weeks) were: Spontaneous remission (SR): 7; Glucantime® (Glu) chemotherapy: 9; Immunotherapy with La, Lv, Lb, Lch amastigotes Tosyl-Lysil-Chloromethyl-Ketone (TLCK) treated and Nonidet P-40 (NP-40) extracted (AS100-1VT): 7. While vaccinating subjects for CL protection, we observed 100% clinical remission of a psoriatic lesion in one subject. In an open trial, 2,770 subjects showed baseline psoriasis area and severity index (PASI) compared with post-treatment values as follows: PASI 100, 23%; PASI 75, 45%; PASI 50, 13%; PASI 10, 9% and <PASI 10, 3% of patients, without serious adverse events. Similar results were obtained with AS100-2 La, Lv, Lb and Lch monovalent vaccines. After treatment with AS100-1VT, subjects with psoriatic arthritis (PsA) decreased in arthritis score, tender joints counts and nail changes; the highest decreased in the PASI 100 group. The vaccines induced cellular immunity with absence of humoral antibody responses. Lymphocyte subsets (LS) in peripheral blood mononuclear cells (PBMC) decrease as PASI increased migrating from the blood to the skin/joints. After vaccine treatment the LS migration is stopped explaining remission of lesions. Purified Leishmania antigenic fractions (AS200) induced linear delayed type hypersensitivity reactions (DTH) in guinea pigs. A DBA-1 mouse collagen induced arthritis (CIA) model with AS200 treatment had the least amount of forepaw inflammation and the lowest arthritis scores, lower than dexamethasone. The vaccines AS100-1VT, AS100-2 and AS200 were not immunosuppressors, but immunomodulators decreasing inflammatory cytokines. The aim of this review is to explain the serendipity discovery of leishmania parasites inducing remission of psoriasis and related diseases, unprecedented discovery in the scientific literature.

A comparison of the molecular biology of trypanosomes and leishmaniae, and its impact on the development of methods for the diagnosis and vaccination of leishmaniasis and Chagas disease

Our main interest have focused on Chagas disease and Leishmaniasis, working in the areas of: 1--The molecular biology of Trypanosomes and Leishmaniae, and 2--The immunology of Chagas disease, cutaneous leishmaniasis and visceral leishmaniasis. In this article we summarize the work realized in the last 20 years in the Immunobiology Laboratory at the IVIC with special emphasis in the development of a vaccine against leishmaniasis that is being currently used in a field trial in human beings of the endemic area of Guatire, Miranda State, Venezuela

Proteínas de la superficie y proteínas excretadas por trypanosoma cruzi y tripanosoma rangeli su importancia en el diagnóstico y la patogenia de la enfermedad de chagas / Surface proteins and excreted proteins by trypanosoma cruzi and trypanosoma rangeli: importance on the diagnosis and pathogenesis of chagas disease

Se aislan y se caracterizan los antigenos de la superficie y los antigenos excretados al medio de cultivo por T.cruzi y T.rangeli, cultivados entre 26§ y 34§C en medios químicamente definidos, donde estos parásitos crecen sin suplemento proteico alguno. T.cruzi muestra 25-30 proteínas en su superficie, siendo las más concentradas las de 107,95,80,77,55,49,47 kDa de peso molecular, no encontrándose variación en el patrón electroforético en SDA-acrimilamida en las distintas cepas, como tampoco a las diversas temperaturas de cultivo. T.rangeli presenta 10-11 proteínas en su superficie, siendo las de mayor concentración a 26§C las correspondientes a 115,106,88,63,50 y 38 kDa, mientras que a 30§C, estas mismas bandas más las correspondientes a 76,57 y 53 kDa predominan sobre las demás. T.cruzi excreta 2 proteínas, una con un peso molecular de 83 kDa y otra cuyo peso molecular abarca un rango de 64 a 76 kDa. La separación de estas proteínas por isoelectroenfoque permite visualizar 12 bandas proteícas con puntos isoeléctricos entre 4,72 y 5,51 con un mayor número de bandas a 34§C que a 26§C, siendo el patrón igual en todas las cepas estudiadas en este trabajo. Al separar las bandas de proteínas onservadas en el isoelectroenfoque madiante cromatografía en DEAE-Sephadex, se observaron 10-15 especies proteícas en cada fracción de la columna, locual permite concluir que el T. cruzi excreta entre 100-150 proteínas de puntos isoeléctricos diferentes. Los sueros de pacientes chagásicos poseen un grupo de anticuerpos que reconocen a las proteínas de superficie de T. cruzi y otros grupos de anticuerpos que reconocen a las proteínas..

Antigénos supresores y protectores de epimastigotes de trypanosoma cruzi expresados a temperaturas diferentes / Supressor and protector antigens of epimastigotes of T.cruzi expressed on different temperatures

Epimastogotes de Trypanosoma cruzi crecido en medios de cultivos que permiten su proliferación entre 26§ y 37§C cambian la estructura de sus proteínas de superficie cuando se eleva la temperatura del cultivo de 30§ a 34§C. Los animales inmunizados con proteínas de superficie de epimastigotes cultivados a 30§C o con sedimentos extraídos con Nonidet P40 y cultivados a 34§C, mostraron parasitemias altas después de la infección con trypomastigotes sanguicolas. En cambio, los animales inmunizados con proteínas de superficie de epimastigotes cultivados a 34§C o con sedimentos de parásitos extraídos con Nonidet P40 y cultivados a 30§C, mostraron parasitemias bajas después de la infección con T.cruzi. La inmunización de ratones con los sedimentos de los parásitos cultivados a 30§C, incubados con tosyl-L-lysina-clorometil-cetona y extraídos con Nonidet P40, produjo la parasitemia menor después de la infección con T.cruzi. así como también ausencia de inmunosupresión y el mayor tiempo de sobrevida, sin evidencias histopatológicas de nidos de amastigotes o infiltrados inflamatorios en el corazón de los animales infectados

Splenocyte membrane changes and inmunosuppression during infecion and reinfection with Trypanosoma cruzi

Antisera to epi- and trypomastigote forms of Trypanosoma cruzi were used to defect trypanosome antigens on the surface of lymphocytes from infected mice. Only the antitrypomastigote serum could recognize antigens expressed transiently on the splenocyte membranes from infected animals. The number or structural configuration of Concanavalin A receptors was similarly affected and a clear correlation was seen between these two types of membrane changes and the immunosuppression to mitogens and SRBC presented by the infected mice. Reinfected animals did not show evidences of trypanosome proliferation in blood or tissues nor trypamastigote antigens on splenocytes, but presented a less intense, transient immunosuppression as measured by responsiveness to mitogens and SRBC, suggesting that the primed immune system can eliminate the new parasite inoculum before the host is immunosuppressed and also that the liberation of strong immunosuppressor trypomastigote antigens induce the new state of suppression

Increase in growth rates and yields by agitation of Trypanosoma cruzi cultures in liquid medium

Trypanosoma cruzi was grown in semi-synthetic medium. Increases in specific growth rate and cell yields were obtained by agitation when compared with stationary cultures. The possible causes for this improvents are briefly discussed